While all eukaryotic cells need a transport system to distribute cargo within the cell, neurons require one with a particularly high degree of spatial and temporal organization. Neurons are highly polarized cells that develop two distinct processes, the axon and dendrites. In order to maintain the complex differential distribution of organelles, cytoskeletal elements and membrane proteins, microtubule-based transport within these two domains must be precisely regulated. Membrane proteins are synthesized in the cell body and must travel to their final destination, sometimes over extremely long distances. This transport occurs via microtubule "tracks", along which the motors kinesin and dynein actively transport cargo, using energy derived from ATP hydrolysis. Microtubule-based transport plays an important role in determining where cargo is delivered. Yet, despite extensive research on kinesins, critical biological questions remain unanswered: What determines the specificity of motor-cargo interactions? How does a motor faithfully deliver its cargo to the correct destination? This proposal focuses on the study of KIF5, also known as conventional kinesin. Its aim is to determine how KIF5 is regulated to transport cargo to the correct destination.